Changes in Blood Parameters After Intramuscular Testosterone Ester Injections - Implications for Anti-Doping

Changes in Blood Parameters After Intramuscular Testosterone Ester Injections : Implications for Anti-Doping / Sara Amalie Solheim, Jakob Mørkeberg, Yvette Dehnes, Ingunn Hullstein, Anders Juul, Emmie N. Upners, Nikolai Baastrup Nordsborg. - (Drug Testing and Analysis (2020) 20 April)

  • PMID: 32307878.
  • DOI: 10.1002/dta.2803


Testosterone treatment stimulates red blood cell production and alters iron homeostasis. Thus, we investigated whether the 'hematological module' of the Athlete Biological Passport (ABP) used by the World Anti-Doping Agency (WADA) can be used to indicate misuse of testosterone. Nineteen eugonadal men received intramuscular injections of either 250 mg Sustanon®, a blend of four testosterone esters, or placebo on Day 0 and 21 in a randomized, placebo-controlled double-blind design. Urine samples and blood samples for the assessment of steroidal and hematological biomarkers of the ABP were collected twice pre-treatment, at least 5 days apart, and on days 1, 3, 5, 10 and 14 post-injections. The steroidal profile was flagged suspicious in all Sustanon®-treated subjects, while the hematological profile was flagged suspicious in 6 out of 9 subjects. When considering both sensitivity and specificity, RET% appeared as the best marker of the hematological module for implying testosterone ester misuse. Atypical blood passport samples were used to select time points for further isotope-ratio mass spectrometry (IRMS) analysis of testosterone and its metabolites in simultaneously collected urine. The RET% and OFF-Score could, in addition to the T/E ratio, help identify suspicious samples for more targeted IRMS testing. The present results demonstrate that unexpected fluctuations in RET% can be indicative of testosterone doping if samples are collected 3-10 days after injection. From an anti-doping perspective, the hematological and steroidal modules of the ABP should complement each other when planning targeted follow-up testing and substantiating likely misuse of testosterone.


20 April 2020
Dehnes, Yvette
Hullstein, Ingunn R.
Juul, Anders
Mørkeberg, Jakob
Nordsborg, Nikolai Baastrup
Solheim, Sara Amalie
Upners, Emmie N.
Adverse Analytical Finding / presence
Other organisations
Anti-Doping Denmark (ADD)
Københavns Universitet - University of Copenhagen (UCPH)
Oslo universitetssykehus HF - Oslo University Hospital
Oslo, Norway: Norwegian Doping Control Laboratory
Analytical aspects
Mass spectrometry analysis
Testing method development
Doping classes
S1. Anabolic Agents
T/E ratio (testosterone / epitestosterone)
Medical terms
Hematocrit levels
Athlete Biological Passport (ABP)
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Date generated
22 April 2020
Date of last modification
7 September 2020
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